罗氏荧光定量试剂盒，FastStart SYBR Green

罗氏荧光定量试剂盒，FastStart SYBR Green
Roche

FastStart™ 通用SYBR® Green预混液(ROX)

FastStart Universal SYBR Green Master (Rox)

astStart^™ Universal SYBR^® Green Master (Rox) is a ready-to-use hot start reaction mix for qPCR and RT-qPCR on all real-time PCR systems requiring normalization with ROX.

SYBR^® Green I is a DNA double-strand-specific dye. During each phase of DNA synthesis, the SYBR^® Green I dye, which is included in the reaction mix, binds to the amplified PCR products. The amplicon can be detected by its fluorescence.

Hot start protocols have been shown to significantly improve the specificity, sensitivity, and yield of PCR. Heat-labile blocking groups on some of the amino acid residues of FastStart^™ Taq DNA Polymerase make the modified enzyme inactive at room temperature (+15 to +25°C). Therefore, there is no elongation during the period when primers can non-specifically bind. FastStart^™ Taq DNA Polymerase is activated by removing the blocking groups at a high temperature (i.e., a pre-incubation step at +95°C).

Application

FastStart^™ Universal SYBR^® Green Master (Rox) can be used for the amplification and detection of any DNA or cDNA target, including those that are GC- or AT-rich.
Combine this master mix with Transcriptor First Strand cDNA Synthesis Kit (Roche) to achieve excellent results in two-step qRT-PCR.
FastStart^™ Universal SYBR^® Green Master (Rox) has been used in qRT-PCR^{[1][3][4][6][8]} and qPCR^[2][5][7][9]

Features and Benefits

• 改善PCR灵敏和特异性。
最大限度减低非特异性扩增产物的形成。

• 避免对qPCR结果的过度预计。
消除可增加量化的SYBR Green I结合量的非特异性扩增产物和引物二聚体。
• 对一系列的DNA或cDNA靶标进行扩增和检测。
扩增最长达500 bp的片段，包括高GC或高AT的。

• 在qPCR制备中节省时间和工作。
免去了降成分混合、优化MaCl₂浓度或进行其他耗时优化优化步骤的需要。

• 防止来自残留污染的假阳性结果。
使用这种含有dUTP的预混液配合尿嘧啶-DNA糖基化酶对来自之前PCR反应的残留DNA污染进行清除。

Components

2x浓度的FastStart Universal SYBR Green Master (Rox)预混液含有FastStart Taq DNA聚合酶、反应缓冲液、核苷酸（dATP, dCTP, dGTP, dUTP）、SYBR Green I以及参考染料。

Quality

功能测试：每个批次都采用三种类型模板对在qPCR中的性能进行测试：高GC模板、高AT模板及长片段模板（约440 bp）。